Fast fabrication process of microfluidic devices based on cyclic olefin copolymer

A new low-cost process for fast fabrication of multilayer microfluidic devices using cyclic olefin copolymer film materials is presented. This novel process consists of the fabrication of microfluidic features by xurography, followed by multilayer lamination via cyclohexane vapor exposure. Exposure time to this solvent and compression time were optimized for bond tensile strength. A three-layer microfluidic chip capable of withstanding back pressures up to 23 MPa was fabricated in less than an hour. The suitability of this fast prototyping method for fabrication of functional UV-transparent microfluidic devices was demonstrated by development and testing of a microfluidic mixer and preparation of a polymer monolithic column within the microfluidic channel

Monolithic sorbents for microscale separations

Over the last decade, the miniaturization of analytical systems has become an increasingly important and interesting research area. Miniaturized systems offer many advantages, including reduced reagent and sample consumption, shorter analysis times, portability and disposability. This dissertation describes novel approaches in this direction, focusing on two areas: the miniaturization of existing column chromatographic systems and the development of microfluidic systems in which the separation is performed in a channel on a microchip. A new type of methacrylate-based monolithic capillary columns for liquid chromatography and capillary electrochromatography were prepared within the confines of fused-silica tubing using Starburst dendrimers to affect porosity. The polyamidoamine (PAMAM) dendrimers were incorporated into a solution of functionalized monomer, cross-linker, solvents, and polymerization initiator. Thermal polymerization, followed by the removal of solvent and dendrimers, produced a continuous rod of polymer with uniform porosity. Different column porosities were obtained by varying the amount of the dendrimer template. The chromatographic performance of these monolithic columns was evaluated using a peptides mixture obtained by tryptic digestion of chicken egg lysozyme. A distinct advantage of polymer monolithic stationary phases over conventional packed chromatographic beds is the ability to prepare them easily and rapidly via free radical polymerization within the channels of a microfluidic device. In this work, continuous polymeric beds were prepared within a channel of three different microchip substrates: glass, poly(dimethylsiloxane) and polycarbonate. The methacrylate-based monolith was cast in-situ via UV-initiated polymerization. The functionalization of the inner wall of the channel with methacryloyl groups enabled the covalent binding of the monolith to the wall. The morphology of the wall-anchored monolith was studied by SEM of chip sections, and by SEM of an extruded segment of non-anchored monolith from a separate chip

Different Stationary Phase Selectivities and Morphologies for Intact Protein Separations

The central dogma of biology proposed that one gene encodes for one protein. We now know that this does not reflect reality. The human body has approximately 20,000 protein-encoding genes; each of these genes can encode more than one protein. Proteins expressed from a single gene can vary in terms of their post-translational modifications, which often regulate their function within the body. Understanding the proteins within our bodies is a key step in understanding the cause, and perhaps the solution, to disease. This is one of the application areas of proteomics, which is defined as the study of all proteins expressed within an organism at a given point in time. The human proteome is incredibly complex. The complexity of biological samples requires a combination of technologies to achieve high resolution and high sensitivity analysis. Despite the significant advances in mass spectrometry, separation techniques are still essential in this field. Liquid chromatography is an indispensable tool by which low-abundant proteins in complex samples can be enriched and separated. However, advances in chromatography are not as readily adapted in proteomics compared to advances in mass spectrometry. Biologists in this field still favour reversed-phase chromatography with fully porous particles. The purpose of this review is to highlight alternative selectivities and stationary phase morphologies that show potential for application in top-down proteomics; the study of intact proteins